Preparation of RNA Samples from Mouse Tissues: Difference between revisions
m Added ball bearing step, added step 6 to transfer to a new 1.5 vial. |
m Added where to dispose of the chloroform in case someone forgets. |
||
| Line 21: | Line 21: | ||
#Centrifuge in a cold eppendorf centrifuge (4C) on maximum for 15 minutes. Sample will separate into a lower, red chloroform phase, an interphase and a colorless upper phase which contains the RNA. | #Centrifuge in a cold eppendorf centrifuge (4C) on maximum for 15 minutes. Sample will separate into a lower, red chloroform phase, an interphase and a colorless upper phase which contains the RNA. | ||
#Add 400 uL of 70% ethanol to a fresh tube. | #Add 400 uL of 70% ethanol to a fresh tube. | ||
#Transfer 400 uL of the upper phase to the ethanol tube and mix by vortexing. | #Transfer 400 uL of the upper phase to the ethanol tube and mix by vortexing. The remaining chloroform in the previous vial should be disposed of in the phenol waste container under the hood. | ||
#Remove 700 uL of mixture (invert briefly to mix before taking sample) and add to a spin column in a collection tube. | #Remove 700 uL of mixture (invert briefly to mix before taking sample) and add to a spin column in a collection tube. | ||
#Spin 15s on max. Discard flow through. Add remaining sample, respin and discard flow through. | #Spin 15s on max. Discard flow through. Add remaining sample, respin and discard flow through. | ||